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. Author manuscript; available in PMC: 2020 Mar 1.
Published in final edited form as: Pharmacol Res. 2019 Jan 2;141:331–342. doi: 10.1016/j.phrs.2019.01.002

Fig. 6. cAMP regulation of BCRP expression and activity in human BeWo trophoblast cells treated with anandamide.

Fig. 6.

A. Effects of AEA on intracellular cAMP levels. BeWo cells were treated (± 10 μM AEA; ± 50 μM forskolin, FOR) for 1 h and collected in 0.1 M HCI. Intracellular concentrations of cAMP were determined by ELISA and normalized to protein concentration. Data are presented as mean ± SE (n = 4). B. 8-Br-cAMP abrogates AEA effects on BCRP mRNA. qPCR was performed on BeWo cells treated with AEA (10 μM), 8-Br-cAMP (1 mM), or both for 24 h. Black and white bars represented samples treated in the absence or presence of AEA, respectively. Data are presented as mean ± SE (n = 4). D. Inhibition of BCRP efflux activity by AEA is cAMP-dependent. BeWo cells were treated with either AEA (10 μM), 8-Br-cAMP (1 mM), or both for 24 h. Cells were then trypsinized and subjected to the Hoechst 33342 transport assay as described in Materials and Methods. Data represent the mean ± SE (n = 3-5). Asterisks (*) represent statistically significant differences (p < 0.05) compared to control cells. Daggers (†) represent statistically significant differences (p < 0.05) compared to AEA-treated cells.